β(1-3)-glucan diagnostic assays

Abstract

Methods of isolating beta (1-3)-glucan or beta (1-3)glucan-containing organisms in a sample, or of detecting the presence of beta (1-3)-glucan or beta (1-3)-glucan-containing organisms in a sample, utilizing binding agents for beta (1-3)-glucan, such as LacCer, GalCer, globotriaosylceramide and asialoganglioside-GM1, are described. Methods of diagnosing fungal infection, by detecting beta (1-3)-glucan or beta (1-3)-glucan-containing organisms, are also described. Antibodies and kits useful in the methods are also disclosed.

What is claimed is: 
     
       1. A method of isolating β(1-3)-glucan from a sample, comprising the steps of: a. contacting a sample containing β(1-3)-glucan with a β(1-3)-glucan binding agent, wherein the β(1-3)-glucan binding agent is a glycosphingolipid, under conditions that are suitable for binding of β(1-3)-glucan to the β(1-3)-glucan binding agent to form primary complexes of β(1-3)-glucan bound to β(1-3)-glucan binding agent;   b. isolating the primary complexes; and   c. isolating β(1-3)-glucan from the primary complexes.   
     
     
       2. The method of claim 1, wherein the β(1-3)-glucan binding agent is coupled to a solid phase. 
     
     
       3. The method of claim 2, wherein the solid phase is selected from the group consisting of: a filter, a membrane, a bead, a particle, an organic resin, a microtiter plate and a slide. 
     
     
       4. The method of claim 1, wherein the β(1-3)-glucan binding agent is selected from the group consisting of: lactosyl ceramide, galactosyl ceramide, globotriaosylceramide and asialoganglioside-GM1. 
     
     
       5. The method of claim 1, wherein the β(1-3)-glucan binding agent is lactosyl ceramide or galactosyl ceramide. 
     
     
       6. A method of detecting the presence of β(1-3)-glucan in a sample, comprising the steps of: a. contacting a sample to be tested for the presence of β(1-3)-glucan with a β(1-3)-glucan binding agent, wherein the β(1-3)-glucan binding agent is a glycosphingolipid, under conditions that are suitable for binding of β(1-3)-glucan, to the β(1-3)-glucan binding agent, wherein primary complexes are formed when β(1-3)-glucan is bound to said β(1-3)-glucan binding agent, and   b. performing an assay to determine if said primary complexes are present,   wherein the presence of primary complexes is indicative of the presence of β(1-3)-glucan in the sample.   
     
     
       7. The method of claim 6, wherein the β(1-3)-glucan binding agent is attached to a solid phase. 
     
     
       8. The method of claim 7, wherein the solid phase is selected from the group consisting of: a filter, a membrane, a bead, a particle, an organic resin, a microtiter plate and a slide. 
     
     
       9. The method of claim 6, wherein the sample is attached to a solid phase. 
     
     
       10. The method of claim 9, wherein the solid phase is selected from the group consisting of: a filter, a membrane, a bead, a particle, an organic resin, a microtiter plate and a slide. 
     
     
       11. The method of claim 6, wherein the β(1-3)-glucan binding agent is selected from the group consisting of: lactosyl ceramide, galactosyl ceramide, globotriaosylceramide and asialoganglioside-GM1. 
     
     
       12. The method of claim 11, wherein the β(1-3)-glucan binding agent is lactosyl ceramide or galactosyl ceramide. 
     
     
       13. The method of claim 6, wherein the sample is from an animal source. 
     
     
       14. The method of claim 13, wherein the sample is a biological fluid. 
     
     
       15. The method of claim 6, wherein the sample is from a plant source. 
     
     
       16. The method of claim 6, wherein the sample is from an environmental source. 
     
     
       17. The method of claim 6, wherein the sample is from a food source. 
     
     
       18. The method of claim 6, wherein the sample is from a fermentation source. 
     
     
       19. The method of claim 6, wherein the assay comprises the use of a detectable label for detection of the primary complexes. 
     
     
       20. The method of claim 19, wherein the detectable label is selected from the group consisting of: radionucleotides, dyes, fluorescent compounds, biotin and streptavidin. 
     
     
       21. The method of claim 6, wherein the assay comprises the use of an antibody for detection of the primary complexes. 
     
     
       22. The method of claim 21, wherein the antibody is an antibody to β(1-3)-glucan. 
     
     
       23. The method of claim 21, wherein the antibody is an antibody to β(1-3)-glucan binding agent. 
     
     
       24. A method of quantifying the amount of β(1-3)-glucan in a sample, comprising the steps of: a. contacting a sample to be tested with a β(1-3)-glucan binding agent, wherein the β(1-3)-glucan binding agent is a glycosphingolipid, under conditions that are suitable for binding of β(1-3)-glucan, to the β(1-3)-glucan binding agent, to form primary complexes of β(1-3)-glucan bound to β(1-3)-glucan binding agent;   b. isolating the primary complexes formed in step (a); and   c. quantifying the primary complexes,   wherein the amount of β(1-3)-glucan in the sample correlates with the amount of primary complexes.   
     
     
       25. A method of quantifying the amount of β(1-3)-glucan in a sample, comprising the steps of: a. contacting a sample to be tested with a β(1-3)-glucan binding agent, wherein the β(1-3)-glucan binding agent is a glycosphingolipid, and a labeled, underivatized, aqueous soluble β(1-3)-glucan, under conditions that are suitable for binding of β(1-3)-glucan, to the β(1-3)-glucan binding agent to form primary complexes of underivatized, aqueous soluble β(1-3)-glucan bound to β(1-3)-glucan binding agent; and   b. determining the amount of primary complexes formed in step (a),   wherein the amount of β(1-3)-glucan in the sample is inversely related to the amount of primary complexes.